CAJ | 학술논문

目的：探讨 miRNA-101、ZESTE 基因增强子同源物2（EZH2）及转化生长因子（TGF）-β1在幼鼠单侧输尿管完全梗阻（CUUO）肾脏中的表达和意义。方法选取30只 SD 雄性幼鼠，周龄（6±1）周，体质量（150±10）g，采用随机数字表法分为 sham 组、CUUO 7 d 组、CUUO 14 d 组，每组10只。分别于梗阻后7 d 及14 d 处死取其梗阻侧肾脏，横断面切开分为均等的上下段2部分，上段部分用于 Western blot 检测肾脏 TGF-β1及 EZH2的表达；下段部分再纵切为均等的2部分，一部分用于实时荧光定量-聚合酶链反应（RT-PCR）检测不同梗阻时间段肾脏 miRNA-101的表达，另一部分用于免疫组织化学及 HE 染色检测肾脏 TGF-β1及 EZH2的定性表达。结果 RT-PCR 结果显示 miRNA-101相对表达量：sham 组是 CUUO 14 d 组的（12.69±1.60）倍， CUUO 7 d组是14 d 组的（3.74±1.24）倍，3组间差异有统计学意义（P ﹤0.05）；Western blot 显示 TGF-β1相对表达量在 CUUO 14 d、7 d 及 sham 组分别为1.14±0.12、1.04±0.14和0.76±0.18；EZH2相对表达量在 CUUO 14 d、7 d 及 sham 组分别为1.04±0.04、0.89±0.03和0.73±0.02；miRNA-101与 EZH2表达呈负相关（r ＝-0.92，P ﹤0.05），与 TGF-β1表达呈负相关（r ＝-0.63，P ﹤0.05），EZH2与 TGF-β1表达呈正相关（r ＝0.67， P ﹤0.05）。不同梗阻时间肾脏 miRNA-101、EZH2及 TGF-β1的表达均具有明显相关性。结论随着梗阻时间的延长，miRNA-101表达下降，EZH2及 TGF-β1表达上升，提示 miRNA-101可能通过调节幼鼠肾脏 EZH2表达影响肾间质纤维化进展。
목적：탐토 miRNA-101、ZESTE 기인증강자동원물2（EZH2）급전화생장인자（TGF）-β1재유서단측수뇨관완전경조（CUUO）신장중적표체화의의。방법선취30지 SD 웅성유서，주령（6±1）주，체질량（150±10）g，채용수궤수자표법분위 sham 조、CUUO 7 d 조、CUUO 14 d 조，매조10지。분별우경조후7 d 급14 d 처사취기경조측신장，횡단면절개분위균등적상하단2부분，상단부분용우 Western blot 검측신장 TGF-β1급 EZH2적표체；하단부분재종절위균등적2부분，일부분용우실시형광정량-취합매련반응（RT-PCR）검측불동경조시간단신장 miRNA-101적표체，령일부분용우면역조직화학급 HE 염색검측신장 TGF-β1급 EZH2적정성표체。결과 RT-PCR 결과현시 miRNA-101상대표체량：sham 조시 CUUO 14 d 조적（12.69±1.60）배， CUUO 7 d조시14 d 조적（3.74±1.24）배，3조간차이유통계학의의（P ﹤0.05）；Western blot 현시 TGF-β1상대표체량재 CUUO 14 d、7 d 급 sham 조분별위1.14±0.12、1.04±0.14화0.76±0.18；EZH2상대표체량재 CUUO 14 d、7 d 급 sham 조분별위1.04±0.04、0.89±0.03화0.73±0.02；miRNA-101여 EZH2표체정부상관（r ＝-0.92，P ﹤0.05），여 TGF-β1표체정부상관（r ＝-0.63，P ﹤0.05），EZH2여 TGF-β1표체정정상관（r ＝0.67， P ﹤0.05）。불동경조시간신장 miRNA-101、EZH2급 TGF-β1적표체균구유명현상관성。결론수착경조시간적연장，miRNA-101표체하강，EZH2급 TGF-β1표체상승，제시 miRNA-101가능통과조절유서신장 EZH2표체영향신간질섬유화진전。
Objective To explore the expressions and significance of miRNA - 101,enhancer of ZESTE homo-log 2(EZH2)and transforming growth factor(TGF)- β1 in the kidneys with complete uniateral ureteral obstruction (CUUO). Methods Thirty male SD rats,(6 ± 1)weeks old,weighted(150 ± 10)g,were divided into sham group, 7 - day group with CUUO and 14 - day group with CUUO by using random number table method,10 rats in each group. The obstructed kidney samples were collected in 7 and 14 days,respectively,for detecting the expression of miRNA -101 by real time - polymerase chain reaction(RT - PCR)and TGF - β1 and EZH2 protein by Western blot,immuno-histochemistry and hematoxylin - eosin staining. Their correlated expressions were analyzed. Results RT - PCR results showed that the expressions of miRNA - 101 in sham group and 7 - day group with CUUO were(12. 69 ± 1. 60)times and(3. 74 ± 1. 24)times which were higher than those of 14 - day group with CUUO,respectively,there was a signifi-cant difference among these 3 groups(P ﹤ 0. 05). The expressions of TGF - β1 and EZH2 proteins were 1. 14 ± 0. 12, 1. 04 ± 0. 14,0. 76 ± 0. 18 and 1. 04 ± 0. 04,0. 89 ± 0. 03,0. 73 ± 0. 02 in 14 - day group with CUUO,7 - day group with CUUO and sham group,respectively. There was a negative correlation between miRNA - 101 with EZH2( r =- 0. 92,P ﹤ 0. 05),and negative correlation with TGF - β1(r = - 0. 63,P ﹤ 0. 05),and positive correlation between EZH2 and TGF - β1(r = 0. 67,P ﹤ 0. 05);the expressions of miRNA - 101,EZH2 and TGF - β1 were associated with each other in obstruction renal in different time periods. Conclusions With the extension of obstruction time, miRNA - 101 expression decreased,EZH2 and TGF - β1 expression increased,evidently,which indicates that the de-velopment of renal interstitial fibrosis may be affected through regulating EZH2 expression of renal obstruction through miRNA - 101 in the young rats.